Title: Transforming Escherichia coli with a Recombinant Plasmid Overview: This is another lab from the Amgen Bruce Wallace biotechnology lab book. In this lab the students will be adding a plasmid from sea anemones into a bacterial colony causing them to fluoresce. This will allow the students to observe (artificial) mutation in person. Opening (10 min) Lab work (70 min) Closing (10 min) Standards (Links): https://riscienceteachers.wikispaces.com/LS3+p1 Objectives: Students will be able to... -create a recombinant gene within a bacterial colony to observe mutation Materials / Preparation Notes: Reagents LIG tube (contains recombinant plasmids) 100 µL of competent cells (HB101) 800 µL of LB broth (sterile) Crushed ice (in a styrofoam cup) Agar plates, sterile 1 LB, 1 LB/amp, 1 LB/amp/ara
Overview: This is another lab from the Amgen Bruce Wallace biotechnology lab book. In this lab the students will be adding a plasmid from sea anemones into a bacterial colony causing them to fluoresce. This will allow the students to observe (artificial) mutation in person.
Opening (10 min)
Lab work (70 min)
Closing (10 min)
Standards (Links):
https://riscienceteachers.wikispaces.com/LS3+p1
Objectives:
Students will be able to...
-create a recombinant gene within a bacterial colony to observe mutation
Materials / Preparation Notes:
Reagents
LIG tube (contains recombinant plasmids)
100 µL of competent cells (HB101)
800 µL of LB broth (sterile)
Crushed ice (in a styrofoam cup)
Agar plates, sterile
1 LB, 1 LB/amp, 1 LB/amp/ara
Equipment and supplies
P-20 micropipette and tips
Disposable 1 mL pipette
42°C water bath
1 pack cell spreaders (shared)
Plastic microfuge tube rack
1.5 mL microfuge tubes
Marking pen
Instruction
Opening: I will begin by briefly reading over the lab and answering any questions the students have.
Activities: The lab procedure is located here: http://www.csun.edu/~jcm68622/biology/subjects/biotechnology/student/Student%20Guide%203rd%20Ed.pdf. It is lab 5, pages 5.1-5.4.
Closing: We will close up the lab by going over the questions located on the last page of lab 5.
Assessment Notes: The assessment for this lab will be the completion of the lab and active participation in the post lab questions.